本文關(guān)鍵詞： SPR 生物傳感器 基因芯片 病原體 膠體金　出處：《重慶醫(yī)科大學(xué)》2006年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的:構(gòu)建基于表面等離子體共振(Surface Plasmon Resonance,SPR)原理的病原體快速檢測(cè)基因芯片。利用SPR傳感器的快速、敏感、高效、不需標(biāo)記及純化等特點(diǎn),建立一套完整的基因芯片系統(tǒng)檢測(cè)技術(shù),使之具有設(shè)備簡(jiǎn)單,靈敏度高,應(yīng)用范圍廣,技術(shù)穩(wěn)定易于掌握等優(yōu)點(diǎn),達(dá)到使基因芯片的檢測(cè)技術(shù)能更適用于臨床、普通實(shí)驗(yàn)室甚至更復(fù)雜環(huán)境的目的。 方法:1.通過(guò)查閱文獻(xiàn)報(bào)道以及臨床調(diào)查,確定檢測(cè)靶病原體。對(duì)各靶病原體的特征性核酸標(biāo)志進(jìn)行確定以及擴(kuò)增引物的設(shè)計(jì)并對(duì)擴(kuò)增條件進(jìn)行優(yōu)化確定;2.設(shè)計(jì)各靶病原體特異性寡核苷酸探針,利用生物信息學(xué)方法對(duì)探針進(jìn)行計(jì)算機(jī)模擬篩選;3.在載玻片表面鋪制具有SPR響應(yīng)的50nm金膜層,并比較了常用兩種不同直徑膠體金溶液金膜鋪制方法的效果;4.利用硫醇化合物表面單分子自組裝層技術(shù)(Self-assembled Monolayer,SAM)固定探針,制備具有SPR響應(yīng)的基因檢測(cè)芯片,并對(duì)自組裝時(shí)間,探針濃度等進(jìn)行優(yōu)化;5.將篩選出的檢測(cè)探針、陽(yáng)性對(duì)照探針以及陰性探針點(diǎn)利用SAM技術(shù)制成雜交芯片,利用酶標(biāo)記化學(xué)發(fā)光法對(duì)各探針的雜交性能進(jìn)行檢測(cè);6.利用SPR檢測(cè)系統(tǒng)對(duì)所構(gòu)建芯片的檢測(cè)性能進(jìn)行了測(cè)試;7.結(jié)合Chelex-100處理臨床樣品進(jìn)行檢測(cè),與常規(guī)臨床檢測(cè)方法比較該芯片的檢測(cè)特性。 結(jié)果: 1.確定了7種臨床常見的感染病原體及臨床少見但重要的
[Abstract]:Objective: to construct surface Plasmon Resonance based on surface plasmon resonance (SPR). Based on the characteristics of SPR sensor such as fast sensitivity, high efficiency, no marking and purification, a set of complete gene chip system detection technology was established. It has the advantages of simple equipment, high sensitivity, wide range of application, stable and easy to grasp and so on. It can make the detection technology of gene chip more suitable for clinical, general laboratory and even more complex environment. Methods 1. The target pathogens were determined by consulting literature reports and clinical investigations. The characteristic nucleic acid markers of each target pathogen and the design of amplification primers were determined and the conditions of amplification were optimized. 2. The specific oligonucleotide probes for each target pathogen were designed, and the probes were screened by computer simulation using bioinformatics. 3. A 50nm gold film with SPR response was prepared on the slide surface, and the results of two kinds of gold film preparation methods of colloidal gold solution with different diameters were compared. 4. Self-assembled Monolayerian SAM (Self-assembled Monolayerian SAM) immobilized probe was developed by using the monolayer layer technique on the surface of mercaptan compounds. A gene detection chip with SPR response was prepared and the self-assembly time and probe concentration were optimized. 5. The hybridization microarray was made by using SAM technique, the detected probe, the positive control probe and the negative probe spot were used to detect the hybridization performance of each probe by using enzyme-labeled chemiluminescence method. 6. The performance of the chip is tested by SPR detection system. 7. The detection characteristics of the chip were compared with the conventional clinical detection method. Results: 1. Seven common clinical pathogens and rare but important clinical pathogens have been identified. 2.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R346

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相關(guān)期刊論文 前2條

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本文編號(hào)：1490513